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University of Ghana http://ugspace.ug.edu.gh UNIVERSITY OF GHANA COLLEGE OF HEALTH SCIENCES OCCURRENCE OF BABESIA / THEILERIA AMONGST HUMANS, CATTLE, AND DOGS AT THE MIDDLE BELT OF GHANA BY BENJAMIN PULLE NIRIWA (10600042) A THESIS SUBMITTED TO THE DEPARTMENT OF MEDICAL MICROBIOLOGY OF THE UNIVERSITY OF GHANA IN PARTIAL FULFILLMENT OF THE RE- QUIREMENT FOR THE AWARD OF A MASTER OF PHILOSOPHY DEGREE IN MEDICAL MICROBIOLOGY JULY, 2018 University of Ghana http://ugspace.ug.edu.gh DECLARATION I hereby declare that except for references to other people’s work, which I have duly acknowl- edged, this work is a result of my own research under the able supervision of Dr. Patience Borkor Tetteh-Quarcoo and Rev. Prof. Patrick Ferdinand Ayeh-Kumi, both of the Department of Medical Microbiology, School of Biomedical and Allied Health Sciences, College of Health Sciences, Uni- versity of Ghana. This work neither in whole nor in part had been submitted for another degree elsewhere. BENJAMIN PULLE NIRIWA (STUDENT) Signed: ……………………………. Date: ……………………………… REV. PROF. PATRICK FERDINAND AYEH-KUMI (SUPERVISOR) Signed: …………………………. Date: ……………………………. DR. PATIENCE BORKOR TETTEH – QUARCOO (SUPERVISOR) Signed: …………………………. Date: ……………………………. i University of Ghana http://ugspace.ug.edu.gh DEDICATION This thesis is first dedicated to God Almighty for His divine protection, guidance, and divine mi- raculous favors. I also dedicate it to my church for their prayers and spiritual support. ii University of Ghana http://ugspace.ug.edu.gh ACKNOWLEDGEMENT I will first of all thank Almighty God for His divine protection, guidance, and divine miraculous favors. He protected me throughout this course, despite all the uncertainties that sometimes come my way. Secondly, I thank my entire family for their understanding and prayers. I am very grateful to the entire staffs of the Medical Microbiology Department for providing us with a supportive environment to learn. I am full of gratitude to my supervisors: Rev. Prof. Patrick Ferdinand Ayeh-Kumi and Dr. Patience Borkor Tetteh-Quarcoo, who were like my God-sent an- gels. I did not only learn academically from them. I specially thank all our knowledgeable God- fearing, loving, and understanding lecturers for the knowledge and all that they have impacted in us. Special thank goes to the Departmental Head, Prof. Kwamena W. C. Sagoe; Dr. Nicholas Dzifa Dayie; The Secretaries, especially Mad. Evelyn Omane; and the D - Block hostel managerees, Mad. Gloria Lariba Akan-Yidi. I express my gratitude to Rev. Dr. Charles Antwi - Boasiako for allowing his thermocycler to be used for the PCR part of this work. I am also grateful to Mr. Robert Aryeeh for teaching me how to practically extract DNA from my samples and do the PCR. Thank you also, Mr. Emmanuel Afutu and Mr. Ernest Akomeah for helping with editing of the thesis. I thank management and staffs of Holy Family Hospital, Techiman (especially Mr. Emmanuel Adu and Anthony Appiah Lab.), and Kintampo Municipal Hospital (especially Alidu Batong Ibrahim, who helped in collection of the smear positive malaria patients’ samples). I appreciate the veteri- nary officers at Techiman (especially Dr. Anthony Osei Tutu, Dr. David Antwi, Mr. Frimpong Mensah Emmanuel, Mr. Michael Acquay, and Mr. Richard Dery) and Kintampo for helping with collection of blood samples from the animal. I also express my appreciation to Mr. David Dakura, the Head of Kintampo Veterinary Office, for his generous support. Alhaji Sumani Sarichi, I am grateful to you for giving me an interpreter to help me collect cattle rearers samples. iii University of Ghana http://ugspace.ug.edu.gh ABSTRACT Background: Babesia/Theileria species are intra-erythrocytic protozoa of the phylum apicom- plexa. The merozoite stage of Babesia/Theileria have diagnostic significance and are found as intracellular inclusions of infected red blood cells. The trophozoite stages appear as ring forms which measure about 1.0 to 5.0µm. These parasites are transmitted by hard ticks and can cause a zoonotic disease known as babesiosis/theileriosis. Human babesiosis/theileriosis are usually asymptomatic except in immuno-compromised people in whom symptoms present like malaria, yet treatment for these diseases can be different. These similarities can increase the possibility of misdiagnosing a patient with malaria when he or she is really suffering from babesiosis/theileriosis or vice versa leading to an inappropriate treatment choice. Ghana is a malaria endemic country; thus, general malaise is usually treated as malaria. Aim: The aim of this study was to investigate the occurrence of Babesia/Theileria species in cattle, dogs and humans at the Middle Belt of Ghana. Methodology: A cross-sectional study involving humans, sick dogs and cattle was undertaken at communities within the Techiman and Kintampo municipalities. Microscopy (Giemsa stained thin smears), serology (RDT) and polymerase chain reaction (PCR) were techniques employed in the study. Whole blood samples were taken from sick cattle, sick dogs and human malaria positive cases (that were negative for rapid diagnostic test). Blood samples of all study subjects were mi- croscopically screened to suspect Babesia/Theileria infection. Suspected samples were subse- quently subjected to PCR amplification and sequenced for specific piroplasm and strain identifi- cation. Results: Out of seventy-one (71) sick dogs, 30 (42.3%) were suspected by microscopy whiles 14 out of the suspected cases (14/30, 46.7%) were amplified by PCR. For sick cattle, 33 (15.9%) were suspected out of the two hundred and seven (207), out of which 20 (20/33, 60.6%) were amplified and subsequently sequenced. The piroplasm identified in the cattle after sequencing was Theileria velifera. Twenty (10.7%) out of the one hundred and eighty-seven (187) Plasmodium-like smear positive cases, were suspected (smear positive but RDT negative) of Babesia/Theileria infection, with 6 (30.0%) amplified and identified as Theileria velifera after successful sequencing. Conclusion: Babesia/Theileria has been found in all the study groups (dogs, cattle and humans). This is the first report of human theileriosis (caused by Theileria velifera) in Ghana. iv University of Ghana http://ugspace.ug.edu.gh TABLE OF CONTENTS DECLARATION............................................................................................................................ i DEDICATION............................................................................................................................... ii ACKNOWLEDGEMENT ........................................................................................................... iii ABSTRACT .................................................................................................................................. iv TABLE OF CONTENTS ............................................................................................................. v LISTS OF FIGURES .................................................................................................................... x LIST OF TABLES ....................................................................................................................... xi ABBREVIATIONS ..................................................................................................................... xii CHAPTER ONE ........................................................................................................................... 1 1.0 INTRODUCTION................................................................................................................... 1 1.1 Background ............................................................................................................................. 1 1.2 Problem statement .................................................................................................................. 2 1.3 Justification ............................................................................................................................. 4 1.4 Main Aim ................................................................................................................................. 5 1.5 Specific objectives ................................................................................................................... 6 1.6 Hypothesis ................................................................................................................................ 6 CHAPTER TWO .......................................................................................................................... 7 2.0 LITERATURE REVIEW ...................................................................................................... 7 2.1 Overview Babesia/Theileria species ...................................................................................... 7 v University of Ghana http://ugspace.ug.edu.gh 2.1.1 Classification, Similarities, and Differences between Babesia and Theileria species ..... 8 2.1.2 Life Cycle and Transmission of Babesia/Theileria species ............................................. 13 2.2 Infection of Humans with Babesia/Theileria Species (Human Babesiosis/Theileriosis) 17 2.2.1 How Babesia/Theileria complicates human malaria diagnosis and treatment ……….18 2.3 Babesia/Theileria species In Cattle (Bovine Babesiosis/Theileriosis) ............................... 20 2.4 Babesia/Theileria species in Dogs (Canine Babesiosis/Theileriosis) ................................. 22 2.5 Pathogenesis, Signs
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